Understanding mechanisms of cross talk between Tuberous sclerosis complex and the Notch signalling pathway.

Mutations in human Tuberous sclerosis complex, TSC-1 and TSC-2 genes cause a serious and incurable genetically inherited disorder, which is associated with tumour-like growths with cell-lineage defects in multiple organs. TSC regulates mTor signalling and tor inhibitors are partially effective in supressing some disease manifestations. There is much interest in identifying additional targets in other pathways that We have identified using the Drosophila model organism, a novel output of TSC acting through the cell fate signal Notch.

The outcomes of TSC loss are context dependent, and, in vivo, in different tissues result in either loss or gain of Notch signalling, the former being independent of Tor activity and the latter being dependent on it. Tor independent loss of Notch signalling depends on the activity of endocytic trafficking regulators. We also observe increased cell death in some tissues occurring after TSC2 loss combined with activation of Notch endocytic trafficking.

These results are important because it suggests that targeting of the Notch pathway might be beneficial alongside Tor inhibition to improve patient outcomes and potentially remove tthe tumour-like growths altogether. The aim of this project is to understand the pathways identified further by mapping the structure/function requirement of TSC1 and TSC2 proteins, for the different Tor-dependent and independent outputs that we have identified.

In this project the student will use Drosophila model genetic organism, cell culture and pluripotent stem cell technology along with CRISPR/CAS9 gene editing to elucidate the mechanism by which this cross talk takes place and determine how TSC-1 or 2 disease mutants located in different domains affect different pathways. By understanding which mutations affect which outputs of the TSC complex, the results will contribute ability to stratify different patient mutants for potential targeted therapies.

Working with the Tuberous Sclerosis Association during a three month CASE placement the student will participate in the construction of patient database of disease life histories, learn different aspects of the charities' functions, including organising scientific and lay meetings, fundraising and writing lay articles the association's magazine and website. This project offers a unique opportunity to combine interdisciplinary research while interacting with and participating in the work of a disease focussed charity.

Objectives 1) How do TSC mutants affect Notch signalling and trafficking in cells? 2) How do different TSC mutants affect Tor-independent cross talk with Notch in vivo? 3) How do TSC mutants and Notch interact in pluripotent stem cell derived human kidney cells.

4) In collaboration with the CASE partner organisation the student will develop a health database linking patient genetics with disease life history, treatment outcomes and disease progression, will assist with organisation of scientific and lay meetings of the Tuberous sclerosis community. This will run in parallel to the research objectives 1-3.